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FFPE DNA extraction Embedded tissue DNA Extraction 100 prep

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$169.00

The FFPE DNA Extraction kit with magnetic beads is fast, high yield, high quality, and reliable.  It also can be used in the automatic DNA extraction machine.

FORMALIN-FIXED, PARAFFIN-EMBEDDED (FFPE) (protocol PDF)

The FFPE DNA Extraction kit with magnetic beads is fast, high yield, high quality, and reliable.  It also can be used in the automatic DNA extraction machine.

Each preparation is based on 200 ul of mineral oil and 200 microliters of Lysis buffer. The ethanol is not included in the kit.

Prepare the curls from FFPE tissue blocks

1. Section FFPE tissue blocks

2. Remove paraffin from the sections with paraffin dissolve buffer

3. Wash twice with 100%, 75%, and 50% ethanol

4. Dry the tissue pellet

  1. Cut sections from FFPE tissue blocks using a microtome.

Note: For miRNA extraction, we recommend using sections of 10 µm or thicker.

1. Collect each section in a 1.5-ml microcentrifuge tube.

2. Preheat a heating block (with lid) or incubator at 50°C.

3. Add 1 mL of Dissolve Clearing Agent, or equivalent (xylene, mineral oil, Limonene, and another solvent) to the section, and vortex.

4. Centrifuge briefly to ensure that all the tissue is submerged in the

5. Heat the sample for 3 minutes at 50°C to melt the Centrifuge the sample at maximum speed for 2 minutes to pellet the tissue.

    • If the sample does not form a tight pellet, centrifuge again for 2 minutes.
    • If a tight pellet still does not form, proceed with caution to the next step.
    • Remove and discard the

Note: If the pellet is loose, leave 50–100 µL of solvent in the tube to avoid removing any tissue pieces. The tissue is usually clear and can be difficult to see.

  1. Add 1 mL of 100% ethanol to the tissue pellet and The tissue should turn opaque.
  2. Centrifuge the sample at maximum speed for 2 minutes.
  3. Remove and discard as much ethanol as possible without disturbing the pellet.
  4. Performa second ethanol wash by repeating step 3a through step 3c to ensure the complete solvent

IMPORTANT!  Omit the second wash when working with small samples as excessive washing can result in sample loss.

Times will vary depending on how much ethanol is present. Dry the pellet using one of the following methods:

  • Use a centrifugal vacuum concentrator with one of the following Digest with Protease
  • Air dry at room temperature for 15–45 minutes.

STOPPING POINT (Optional) The dried samples can be stored at room temperature for up to 72 hours.

Add Protease Solution (see Table 5) to each sample according to the following table.

  1. Gently flick the tube to mix and to immerse the

If the tissue sticks to the sides of the tube, use a pipet tip to push the tissue into the solution or centrifuge briefly to immerse the tissue in the solution.

  1. Incubate at 55°C for 1 hour or longer, then centrifuge briefly to collect any condensation droplets.

Note: If you are using an incubator, use a 4-way microtube rack to allow homogeneous incubation of the samples.

Incubate at 90°C for 1 hour, then centrifuge briefly to collect any condensation droplets. Note: Ensure that tubes are tightly capped. Tube caps may pop open during the incubation. Note: For automated isolation, set up the processing plates during the incubation.

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